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Multiplex 3D nanoscopy resolves nanoarchitecture of human immunodeficiency virus

Resolving the nanoscale organization of viral and host proteins is important to understanding virion assembly and infectivity. Here, we present a robust framework for multiplexed optical 3D super-resolution microscopy of human immunodeficiency virus type 1 (HIV-1) particles using minimal fluorescenc...

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Bibliographic Details
Main Authors: Hacke, Moritz (Author) , Kaplan, Charlotte (Author) , Laketa, Vibor (Author) , Heilemann, Mike (Author) , Müller, Barbara (Author) , Kräusslich, Hans-Georg (Author)
Format: Article (Journal)
Language:English
Published: 20 January 2026
In: ACS nano
Year: 2026, Volume: 20, Issue: 2, Pages: 1911-1927
ISSN:1936-086X
DOI:10.1021/acsnano.5c12222
Online Access:Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1021/acsnano.5c12222
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Author Notes:Moritz Hacke, Charlotte Kaplan, Vibor Laketa, Mike Heilemann, Barbara Müller, and Hans-Georg Kräusslich
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Summary:Resolving the nanoscale organization of viral and host proteins is important to understanding virion assembly and infectivity. Here, we present a robust framework for multiplexed optical 3D super-resolution microscopy of human immunodeficiency virus type 1 (HIV-1) particles using minimal fluorescence photon flux (MINFLUX) nanoscopy and DNA point accumulation for imaging in nanoscale topography (DNA-PAINT), achieving isotropic localization precision below 10 nm for five target proteins. First, we assessed linkage errors introduced by different labeling strategies by employing the HIV-1 matrix layer as a reference structure. We then extended the approach to display five viral and host proteins and mapped the spatial organization of tetraspanin proteins CD9 and CD81 in single virus-like particles. For accurate visualization and quantitation of multicolor 3D MINFLUX imaging data, we developed the analysis workflow and software matFLUX. The approach presented here enables high-resolution spatial mapping of protein components within individual virus particles and is generally applicable to the study of nanoscale architectures in 3D.
Item Description:Online veröffentlicht: 14. Dezember 2025
Gesehen am 09.02.2026
Physical Description:Online Resource
ISSN:1936-086X
DOI:10.1021/acsnano.5c12222

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